动植物细胞工程.pptxVIP

4、动物细胞与组织培养;4.1 动物细胞特点;4.2 动物细胞培养定义;类型;4.3 发展历史;4.4 生长特性;Figure?4-2?The fibroblast.????(A) Phase-contrast micrograph of fibroblasts in culture. (B) Drawings of a living fibroblastlike cell in the transparent tail of a tadpole, showing the changes in its shape and position on successive days. Note that while fibroblasts flatten out in culture, they can have more complex, process-bearing morphologies in tissues. (A, courtesy of Daniel Zicha; B, redrawn from E. Clark, Am. J. Anat. 13:351-379, 1912.);常用术语;4.5 体外培养细胞基本技术;4.5.1 体外培养特点;Animal cells are more difficult to culture than microorganisms because they require many more nutrients and typically grow only when attached to specially coated surfaces. Despite these difficulties, various types of animal cells, including both undifferentiated and differentiated ones, can be cultured successfully. ;4.5.2 体外培养工具;Cells/tissues grown in culture long period of time BASIC STERILE TECHNIQUE ? Work environment and surface ? Plasticware and glassware ? Handling techniques ? Sterilization of solutions for maintenance growth or treatment;WORK ENVIRONMENT AND SURFACE Laminar flow hood Relatively enclosed space Little traffic flowing past work space Confined space can be easily cleaned and maintained Must have annual checkup of HEPA filters Should have internal outlets for electricity, vacuum and gas Frequently outfitted with UV light Prevent contamination by daily scrub with 70% ethanol Use closed flask attached to vacuum for spent medium Separate sterile room All incoming air circulated through HEPA filters ;PLASTICWARE AND GLASSWARE Pipettes, bottles, flasks, petri dishes ? Plastic -- better attachment and cell growth of monolayer cell culture -- more expensive than reusable glass -- suitable for storage at 4oC ? Glass -- Can withstand temperatures 0oC use for maintaining frozen stocks -- Steam sterilize by autoclaving * relies on steam pressure and high heat * use fast-dry cycle to dry condensate * use “wet” cyc